A Method to Investigate a Metabolic Process in a Single Neuron and Its Utilization in the Study of Fast Axonal Transport of Acetylcholine in a Cholinergic Neuron of Aplysia

  • Hiroyuki Koike
  • Yoshitomo Umitsu
  • Hiroko Matsumoto


Intracellular recording techniques have been used successfully to analyze the electrical activities of specific single neurons among numerous heterogeneous populations of cells in the central nervous system. We have pursued development of a technique for studying the metabolic processes of single neurons by intracellular injection of radioactive substances via double-barreled Pyrex® capillary micropipettes (Koike et al., 1972; Koike and Tsuda, 1979, 1980; Koike and Matsumoto, 1985). This simple idea met several technical difficulties. (1) How could sufficient amounts of radioactive markers be applied into the soma of a single neuron? (2) How could we know whether all the materials injected were utilized for the metabolism in the neuron or not? (3) How could we eliminate inherent artifacts for this method such as leakage from the cell and reuptake of the leakage into nearby cells as well as intracellular diffusion of the injected substance. Now most of these difficulties have been overcome except for an inherent problem of time-consuming analyses resulting from single injections.


Axonal Transport Cholinergic Neuron Radioactive Substance Diffusion Curve Intracellular Injection 
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  1. Allen, R. D., Metuzals, J., Tasaki, I., Brady, S. T., and Gilbert, S. P., 1982, Fast axonal transport in squid giant axon, Science 218: 1127–1129.PubMedCrossRefGoogle Scholar
  2. Banks, P., Mayor, D., Mitchell, M., and Tomlinson, D., 1971, Studies on the translocation of noradrenalinecontaining vesicles in post ganglionic sympathetic neurones in vitro. Inhibition of movement by colchicine and vinblastine and evidence for the involvement of axonal microtubules, J. Physiol. (Gond.) 216: 625639.Google Scholar
  3. Bridgman, P. C., Kacher, B., and Reese, T. S., 1986, The structure of cytoplasm in directly frozen cultured cells. II. Cytoplasmic domains associated with organelle movements, J. Cell Biol. 102: 1510–1521.PubMedCrossRefGoogle Scholar
  4. Dahlström, A., 1968, Observations on the accumulation of noradrenaline in the proximal and distal parts of peripheral adrenergic nerves after compression, J. Anat. 99: 677–689.Google Scholar
  5. Goldman, J. E., Kim, K. S., and Schwartz, J. H., 1980, Axonal transport of [3H]serotonin in an identified neuron of Aplysia californica, J. Cell Biol. 70: 304–318.Google Scholar
  6. Koike, H., 1979, A new device for controlled intracellular injection using pneumatic pressure, Integr. Control Funct. Brain 2: 39–41.Google Scholar
  7. Koike, H., 1983, Transmitter specific axonal transport of acetylcholine in a neuron of Aplysia: Colchicine resistant and cytochalasin sensitive, Proc. Int. Union Physiol. Soc. 15: 297.Google Scholar
  8. Koike, H., 1984, Evidence of axonal transport of vesicular acetylcholine in a cholinergic neuron of Aplysia, Neurosci. Lett. Suppl. 17: S3.Google Scholar
  9. Koike, H., and Matsumoto, H., 1985, Fast axonal transport of membrane protein and intra-axonal diffusion of free leucine in a neuron of Aplysia, Neurosci. Res. 2: 281–285.PubMedCrossRefGoogle Scholar
  10. Koike, H., and Nagata, Y., 1979, Intra-axonal diffusion of [3H]acetylcholine and [3H]gamma-aminobutyric acid in a neurone of Aplysia, J. Physiol. (Lond.) 295: 397–417.Google Scholar
  11. Koike, H., and Tsuda, K., 1979, Intracellular acetylcholine synthesis and GABA synthesis in some crustacean neurons, in: Neurobiology of Chemical Transmission ( M. Otsuka and Z. Hall, eds.), John Wiley und Sons, New York, pp. 65–76.Google Scholar
  12. Koike, H., and Tsuda, K., 1980, Cellular synthesis and axonal transport of gamma-aminobutyric acid in a photoreceptor cell of the barnacle, J. Physiol. (Lond.) 305:125–138.Google Scholar
  13. Koike, H., Eisenstadt, M., and Schwartz, J. H., 1972, Axonal transport of newly synthesized acetylcholine in an identified neuron of Aplysia, Brain Res. 37: 152–159.PubMedCrossRefGoogle Scholar
  14. Koike, H., Kandel, E. R., and Schwartz, J. H., 1974, Synaptic release of radioactivity after intrasomatic injection of 3H-choline into an identified cholinergic intemeuron of Aplysia californica, J. Neurophysiol. 37: 815–827.Google Scholar
  15. Koike, H., Umitsu, Y., and Matsumoto, H., 1985, Fast axonal transport of membrane protein and acetylcholine in an Aplysia cholinergic neuron: Its modification by local colchicine perfusion or lowering temperature, J. Physiol. Soc. Jpn. 47: 375.Google Scholar
  16. McCaman, R. E., Weinreich, D., and Borys, H., 1973, Endogenous levels of acetylcholine and choline in individual neurons of Aplysia, J. Neurochem. 21: 473–476.PubMedCrossRefGoogle Scholar
  17. McLean, D. B., and Lewis, S. F., 1984, Axoplasmic transport of somatostatin and substance P in the vagus nerve of the rat, guinea pig, and cat, Brain Res. 307: 135–145.CrossRefGoogle Scholar
  18. Price, C. H., and McAdoo, D. J., 1981, Localization of axonally transported [3H]glycine in vesicles of identified neurons, Brain Res. 219: 307–315.PubMedCrossRefGoogle Scholar
  19. Vale, R. D., Reese, T. S., and Sheetz, M. P., 1985a, Identification of a novel force-generating protein, kinesin, involved in microtubule-based motility, Cell 42: 39–50.Google Scholar
  20. Vale, R. D., Schnapp, B. J., Mitchison, T., Steuer, E., Reese, T. S., and Sheetz, M. P., 19856, Different axoplasmic proteins generate movement in opposite directions along microtubules in vitro, Cell 43: 623–632.Google Scholar

Copyright information

© Springer Science+Business Media New York 1988

Authors and Affiliations

  • Hiroyuki Koike
    • 1
  • Yoshitomo Umitsu
    • 1
  • Hiroko Matsumoto
    • 1
  1. 1.Department of NeurophysiologyTokyo Metropolitan Institute for Neurosciences, Fuchu CityTokyo 183Japan

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